Liu Nan-Nan, Guo Bai-Hui, Wang Lei, Wang Xiao-Xi, Wang Xin, Meng Yan-Li, Tang Gui-Xin, Wang Wei-Ming
College of Traditional Chinese Medicine, Heilongjiang Academy of Chinese Medicine Sciences, Harbin, China.
Advanced Microscopy and Instrumentation Research Center, Harbin Institute of Technology, Harbin, China.
Front Pharmacol. 2025 Mar 26;16:1397543. doi: 10.3389/fphar.2025.1397543. eCollection 2025.
Ophiopogonanone B is a potent component of Qinbai Qingfei-concentrated pills (Qinbai), a new traditional Chinese medicine developed by our hospital for the treatment of pneumonia in children. We aim to study how ophiopogonanone B influences the expression of transient receptor potential anchor protein 1 (TRPA1), substance P (SP), and calcitonin gene-related peptide (CGRP) to treat coughing in -infected mice.
Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to detect ophiopogonanone B. Molecular docking of ophiopogonanone B with TRPA1 was performed using Autodock Vina 1.1.2, and subsequent visualization and analysis of docking outcomes were facilitated using Pymol 2.1 and Discovery Studio. For the evaluation of the pathological structure and morphology, lung tissue sections from mice were prepared for animal experiments and subjected to hematoxylin-eosin (HE) and Masson staining. The impact of ophiopogonanone B on the protein and mRNA expression levels of TRPA1, SP, and CGRP in mouse lung tissue was assessed using immunohistochemistry and real-time polymerase chain reaction (RT-PCR).
The samples acquired through Biacore fishing, which were identified and analyzed by UPLC-Q-TOF-MS, confirmed the presence of ophiopogonanone B. This compound exhibited robust and specific binding affinity for TRPA1. Histological staining using HE and Masson techniques revealed that the lung tissue morphology and structure in the ophiopogonanone B-treated group closely mirrored those observed in the blank group. Subsequent immunohistochemistry and RT-PCR revealed a significant reduction ( < 0.01 or < 0.05) in the proteins and mRNA expression levels of TRPA1, SP, and CGRP in the lung tissue of mice treated with high and medium doses of ophiopogonanone B.
By decreasing the expression of TRPA1, SP, and CGRP in the lung tissues of mice afflicted with coughing due to infection, ophiopogonanone B effectively alleviated post-infection cough symptoms.
麦冬双黄酮B是我院研制的用于治疗小儿肺炎的新型中药——芩百清肺浓缩丸（芩百）的一种有效成分。我们旨在研究麦冬双黄酮B如何影响瞬时受体电位锚蛋白1（TRPA1）、P物质（SP）和降钙素基因相关肽（CGRP）的表达，以治疗感染后咳嗽的小鼠。
采用超高效液相色谱-四极杆飞行时间质谱联用技术（UPLC-Q-TOF-MS）检测麦冬双黄酮B。使用Autodock Vina 1.1.2对麦冬双黄酮B与TRPA1进行分子对接，并使用Pymol 2.1和Discovery Studio对对接结果进行可视化和分析。为评估病理结构和形态，制备小鼠肺组织切片用于动物实验，并进行苏木精-伊红（HE）和Masson染色。采用免疫组织化学和实时聚合酶链反应（RT-PCR）评估麦冬双黄酮B对小鼠肺组织中TRPA1、SP和CGRP蛋白及mRNA表达水平的影响。
通过Biacore筛选获得的样品经UPLC-Q-TOF-MS鉴定和分析，证实了麦冬双黄酮B的存在。该化合物对TRPA1表现出强大且特异性的结合亲和力。使用HE和Masson技术进行的组织学染色显示，麦冬双黄酮B治疗组的肺组织形态和结构与空白组密切相似。随后的免疫组织化学和RT-PCR显示，高剂量和中剂量麦冬双黄酮B治疗的小鼠肺组织中，TRPA1、SP和CGRP的蛋白及mRNA表达水平显著降低（<0.01或<0.05）。
麦冬双黄酮B通过降低感染后咳嗽小鼠肺组织中TRPA1、SP和CGRP的表达，有效减轻了感染后咳嗽症状。
